Summary
Insulin secretion depends critically on glucose metabolism. We investigated whether a rapid viability test could be established for assessing glucose metabolism in insulin secreting cells. The MTT (C,N-diphenyl-N′-4,5-dimethyl thiazol 2 yl tetrazolium bromide) colorimetric assay (reduction of tetrazolium salt to formazan) was applied to rat islets and rat insulinoma cell lines. It was found that the rate of formazan production correlated with glucose oxidation and glucose utilization at glucose concentrations which also stimulated insulin secretion. In differentiated insulinoma INS-1 cells, salt reduction paralleled the insulin release, at glucose concentrations of up to 8.3 mmol/l. The glucose-induced formazan production in INS-1 cells and islets was abolished by exposure to the Beta-cell cytotoxic agents, streptozotocin or alloxan. The MTT assay thus provides a convenient tool for the rapid assessment of Beta-cell metabolism and viability.
Article PDF
References
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55–63
Heo DS, Park JG, Hata K, Day R, Heberman RB, Whiteside TL (1990) Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. Cancer Res 50: 3681–3690
Meglasson MD, Matschinsky FM (1986) Pancreatic islet glucose metabolism and regulation of insulin secretion. Diab Metab Rev 2: 163–214
Asfari M, Janjic D, Meda P, Li G, Halban PA, Wollheim CB (1992) Establishment of 2-mercaptoethanol dependent differentiated insulin secreting cell lines. Endocrinology 130: 167–178
Chick WL, Warren S, Chute RN, Like AA, Lauris V, Kitchen KC (1977) A transplantable insulinoma in the rat. Proc Natl Acad Sci USA 74: 628–632
Halban PA, Praz GA, Wollheim CB (1983) Abnormal glucose metabolism accompanies failure of glucose to stimulate insulin release from a rat pancreatic cell line (RIN m5F). Biochem J 212: 439–443
Malaisse W (1990) Defects of signal transduction in a tumoral islet cell line. In: Okamoto H (ed) Molecular biology of the islets of Langerhans. University Press, Cambridge, pp 315–339
Eizirik D, Sandler S (1989) Function and metabolism of pancreaticβ-cells maintained in culture following experimental induced damage. Pharmacol Toxicol 65: 163–168
Slater TF, Sawyer B, Sträuli U (1963) Studies on succinate-tetrazolium reductase systems. (III) Points of coupling of four different tetrazolium salts. Biochim Biophys Acta 77: 383–393
Hope BT, Michael GJ, Knigge KM, Vincent SR (1991) Neuronal NADPH diaphorase is a nitric oxide synthase. Proc Natl Acad Sci USA 88: 2811–2814
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Janjic, D., Wollheim, C.B. Islet cell metabolism is reflected by the MTT (tetrazolium) colorimetric assay. Diabetologia 35, 482–485 (1992). https://doi.org/10.1007/BF02342448
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF02342448